Background of TP53 / p53 antibody

The 393 amino acid tumor suppressor phosphoprotein p53 plays a major role in the cellular response to DNA damage and other genomic aberrations. The activation of p53 can lead to either cell cycle arrest and DNA repair, or apoptosis. p53 is phosphorylated at multiple sites in vivo and by several different protein kinases in vitro. p53 can apparently be phosphorylated by ATM, ATR, and DNA-PK at Ser15; the phosphorylation impairs the ability of MDM2 to bind p53, promoting both the accumulation and functional activation of p53 in response to DNA damage. The level of p53 protein is low in normal cells but increases following DNA damage or cellular stress. Mutations in the p53 gene are found in more than 50% of human cancers. p53 acts to arrest the cell cycle or induce apoptosis by regulating the transcription of specific genes. p53 can also initiate apoptosis through a direct signaling pathway. The p53 tumor suppressor gene encodes a transcription factor that contributes to several cellular activities that include apoptosis, transient growth arrest, and sustained growth arrest or senescence. Mutations within the p53 gene are found in about half of all human cancers. In cells that are functioning normally the MDM2 protein binds to p53 and maintains p53 at low levels by increasing its susceptibility to proteolysis by the 26S proteosome. A cell that undergoes stress loses the ability of MDM2 to bind to p53 and as a result p53 levels increase which then leads to cell cycle arrest or apoptosis. p53 induced cell cycle arrest or apoptosis can be achieved through transcriptional regulation of several genes including the cell cycle inhibitor p21, DNA repair gene GADD45, and the apoptotic inducer Bax. Besides MDM2 inactivation, p53 can also be functionally inactivated by mutation or binding to DNA tumor virus encoded proteins, such as SV40 large T antigen, Adenovirus E1B and papilloma virus E6 proteins.

Detection of phosphorylated p53 with western blotting using anti-p53 Ser392 monoclonal antibody (AM20020AF-N). After blotting Raji cell crude extract, PVDF membranes were treated with phophatase. Clear 53kD bands were shown using AM20020AF-N before the treatment. After the treatment, the 53kD bands were completely disappeared. Whereas, anti-p53 monoclonal antibody (DO-1) strongly reacted with the treated membrane as well as non treated membrane.

Figure 1. Western blot of rat brain nuclear fraction lysate showing specific immunolabeling of the ~53k p53 phosphorylated at Ser392 (Control). The phosphospecificity of this labeling is shown in the second lane (lambda-phosphatase). The blot is identical to the control except that it was incubated in lambda-Ptase (1200 units for 30 min) before being exposed to the phospho Ser392 p53 antibody. The immunolabeling is completely eliminated by treatment with lambda-Ptase.

Formalin-fixed and paraffin-embedded human cancer tissue reacted with the primary antibody, which was peroxidase-conjugated to the secondary antibody, followed by AEC staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated.

Dot blot analysis of anti-p53 pThr55 Pab (Cat#AP12907PU-N) on nitrocellulose membrane. 50ng of Phospho-peptide or Non Phospho-peptide per dot were adsorbed. Antibody working concentrations are 0.5ug per ml.

Confocal microscopy of human HeLa cells using anti-p53 (BP53-12; FITC) (GTX78241). The expression of p53 protein was enhanced by intercalating reagent. Cells were fixed and permeabilized before incubation with the p53-FITC MAb.

Confocal microscopy of human HeLa cells using anti-p53 (BP53-12; FITC) (GTX78241). The expression of p53 protein was enhanced by intercalating reagent. Cells were fixed and permeabilized before incubation with the p53-FITC MAb.

Identification of TP53 (phospho-S46) monoclonal antibody, clone 36 ( Cat # MAB0659 ) by Western blotting. Using crude cell extracts of MOLT-4 untreated ( left lanes ) and treated with adriamycin for 24 h ( right lanes ). The left panel is the result with our product and right panel is the one obtained withthe product of our competitor. The lower panel is whole TP53 protein identified by omnipotent anti-TP53 antibody.

Western Blot: p53 [phospho Ser392] Antibody [NB200-156] - rat brain nuclear fraction lysate showing specific immunolabeling of the ~53k p53 phosphorylated Rat Ser392 (Control). The phosphospecificity of this labeling is shown in the second lane (lambda-phosphatase: ?-Ptase). The blot is identical to the control except that it was incubated in ?-Ptase (1200 units for 30 min) before being exposed to the phospho Ser392 p53 antibody. The immunolabeling is completely eliminated by treatment with ?-Ptase.

Formalin-fixed and paraffin-embedded human cancer tissue reacted with TP53 (phospho S20) polyclonal antibody ( Cat # PAB0561 ) which was peroxidase-conjugated to the secondary antibody followed by AEC staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. BC = breast carcinoma.

Formalin-fixed and paraffin-embedded human cancer tissue reacted with TP53 (phospho S315) polyclonal antibody ( Cat # PAB0562 ) which was peroxidase-conjugated to the secondary antibody followed by AEC staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. HC = hepatocarcinoma.

Formalin-fixed and paraffin-embedded human cancer tissue reacted with TP53 (phospho T18) polyclonal antibody ( Cat # PAB0568 ) which was peroxidase-conjugated to the secondary antibody followed by AEC staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. HC = hepatocarcinoma.

ELISA was performed using a serial dilution of TP53 polyclonal antibody ( Cat # PAB14128 ), crude serum and flow through in antigen coated wells. By plotting the absorbance against the antibody dilution, the titer of the purified antibody was estimated to be 1 : 4,000.

ChIP assays were performed using the U-2 OS ( human osteosarcoma cell line ), TP53 polyclonal antibody ( Cat # PAB14128 ) and optimized PCR primer sets for PCR.Chromatin sheared from 1x106 cells and 2 µg of antibody against TP53, or beads only were used per ChIP experiment.Figure shows the recovery expressed as a % of input ( the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis ).Upper : recovery by TP53 of the TP21 promoter, a known target for TP53 ( 1 ), or beads only.Recovery of the TP21 promoter by the TP53 antibody is evident based on fluorescent qPCR analysis of immunoprecipitated DNA.Right : recovery of the GAPDH promoter ( used as a negative control ) by TP53 or beads only.

Formalin-fixed and paraffin-embedded human lung carcinoma tissue reacted with TP53 polyclonal antibody ( Cat # PAB4850 ) , which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry ; clinical relevance has not been evaluated.

Western blot of rat brain nuclear fraction lysate showing specific immunolabeling of the ~53k TP53 phosphorylated at Ser392 (Control).The phosphospecificity of this labeling is shown in the second lane (lambda-phosphatase: l-Ptase).The blot is identical to the control except that it was incubated in l-Ptase (1200 units for 30 min) before being exposed to the Phospho-TP53 S392 polyclonal antibody ( Cat # PAB9604 ).The immunolabeling is completely eliminated by treatment with l-Ptase.

Detection of activated p53 Recombinant C-terminal fragment of p53 was incubated with Casein Kinase II in the absence (1) or presence (2) of ATP. Proteins were separarted by SDS-PAGE and transferred to a PVDF membrane. The immunoblot was probed with Monoclonal p53 antibody (clone 9F4) (0.5 µg/ ml) for 1h at RT and developed by ECL (exp. time: 30 sec).

Formalin-fixed and paraffin-embedded human cancer tissue reacted with the primary antibody, which was peroxidase-conjugated to the secondary antibody, followed by AEC staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated.

Figure 2. Formalin-fixed and paraffin-embedded human cancer tissue reacted with the primary antibody, which was peroxidase-conjugated to the secondary antibody, followed by AEC staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated.

Western blot analysis of acetylated p53 (Lys382) expression using AM26538AF-S. MCF7 cells in culture were treated with actinomycin D at 5 nM for the indicated periods and the cell extracts were analyzed by western blotting with AM26538AF-S and omnipotent anti-p53 antibody (DO-1). Acetylation of p53 at Lys382 was induced by the DNA damaging treatment.

Western blot analysis of acetylated p53 (Lys120) expression in HCT116. Samples: Crude cell extracts of HCT116 control (left lanes) and treated with siRNA to knockdown the expression of a Tip60 interacting protein. Total p53 was immunoprecipitated with omnipotent anti-p53 monoclonal antibody (Clone DO-1) from the crude extracts and analyzed by western blotting with anti-p53 antibody (FL393) (upper panel) or AM26539AF-S (middle panel). The lower panel shows total p53 level analysed by western blotting with clone DO-1.

Immunohistochemical analysis of p53 (pS15) staining in human breast cancer formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). The section was then incubated with the antibody at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

Immunofluorescent analysis of p53 (pS15) staining in PC12 cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with the primary antibody in 3% BSA-PBS and incubated overnight at 4 C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight 594-conjugated secondary antibody (red) in PBS at room temperature in the dark. DAPI was used to stain the cell nuclei (blue).

Immunohistochemical analysis of p53 (pS315) staining in human breast cancer formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). The section was then incubated with the antibody at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

Immunofluorescent analysis of p53 (pS315) staining in A431 cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with the primary antibody in 3% BSA-PBS and incubated overnight at 4 C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight 594-conjugated secondary antibody (red) in PBS at room temperature in the dark. DAPI was used to stain the cell nuclei (blue).

Immunohistochemical analysis of p53 staining in human breast cancer formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). The section was then incubated with the antibody at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

Immunofluorescent analysis of p53 staining in HeLa cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with the primary antibody in 3% BSA-PBS and incubated overnight at 4 C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight 594-conjugated secondary antibody (red) in PBS at room temperature in the dark. DAPI was used to stain the cell nuclei (blue).

Immunohistochemical analysis of p53 staining in human lung cancer formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). The section was then incubated with the antibody at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

Immunofluorescent analysis of p53 staining in HepG2 cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with the primary antibody in 3% BSA-PBS and incubated overnight at 4 C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight 594-conjugated secondary antibody (red) in PBS at room temperature in the dark. DAPI was used to stain the cell nuclei (blue).

Immunohistochemical analysis of p53 staining in human lung cancer formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). The section was then incubated with the antibody at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

Immunofluorescent analysis of p53 staining in HeLa cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with the primary antibody in 3% BSA-PBS and incubated overnight at 4 C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight 594-conjugated secondary antibody (red) in PBS at room temperature in the dark. DAPI was used to stain the cell nuclei (blue).

Immunohistochemical analysis of p53 (pS37) staining in human breast cancer formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). The section was then incubated with the antibody at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

Formalin-fixed and paraffin embedded human lung carcinoma labeled with Anti-P53(wt-p53)Polyclonal Antibody, Unconjugated (bs-2090R) at 1:200 followed by conjugation to the secondary antibody and DAB staining.

Western blot of rat brain nuclear fraction lysate showing specific immunolabeling of the ~53k using p53 (phospho Ser392) antibody (GTX23257) p53 phosphorylated at Ser392 (Control). The phosphospecificity of this labeling is shown in the second lane (lambda-phosphatase: λ-Ptase). The blot is identical to the control except that it was incubated in λ-Ptase (1200 units for 30 min) before being exposed to the phospho Ser392 p53 antibody. The immunolabeling is completely eliminated by treatment with λ-Ptase.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY TP53 (RC200003, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-TP53.

OriGene overexpression protein microarray chip was immunostained with UltraMAB anti-TP53 mouse monoclonal antibody (UM800098). The positive reactive proteins are highlighted with two red arrows in the enlarged subarray. All the positive controls spotted in this subarray are also labeled for clarification.(1:100)

Immunohistochemical analysis of p53 (pS392) staining in human lung cancer formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). The section was then incubated with the antibody at room temperature and detected using an HRP conjugad compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

Immunofluorescent analysis of p53 (pS392) staining in HeLa cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with the primary antibody in 3% BSA-PBS and incubated overnight at 4 deg C in a hidified chamber. Cells were washed with PBST and incubated with a DyLight 594-conjugated secondary antibody (red) in PBS at room temperature in the dark. DAPI was used to stain the cell nuclei (blue).

Immunohistochemical analysis of p53 (AcK319) staining in human lung cancer formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). The section was then incubated with the antibody at room temperature and detected using an HRP conjugad compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

Immunofluorescent analysis of p53 (AcK319) staining in HeLa cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with the primary antibody in 3% BSA-PBS and incubated overnight at 4 deg C in a hidified chamber. Cells were washed with PBST and incubated with a DyLight 594-conjugated secondary antibody (red) in PBS at room temperature in the dark. DAPI was used to stain the cell nuclei (blue).

Immunohistochemical analysis of p53 (AcK381) staining in human lung cancer formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). The section was then incubated with the antibody at room temperature and detected using an HRP conjugad compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

Immunofluorescent analysis of p53 (AcK381) staining in HeLa cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with the primary antibody in 3% BSA-PBS and incubated overnight at 4 deg C in a hidified chamber. Cells were washed with PBST and incubated with a DyLight 594-conjugated secondary antibody (red) in PBS at room temperature in the dark. DAPI was used to stain the cell nuclei (blue).

Immunohistochemical analysis of p53 staining in human lung cancer formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). The section was then incubated with the antibody at room temperature and detected using an HRP conjugad compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

Immunofluorescent analysis of p53 staining in HeLa cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with the primary antibody in 3% BSA-PBS and incubated overnight at 4 deg C in a hidified chamber. Cells were washed with PBST and incubated with a DyLight 594-conjugated secondary antibody (red) in PBS at room temperature in the dark. DAPI was used to stain the cell nuclei (blue).

p53 Antibody (T55) (Cat. #TA302001)immunohistochemistry analysis in formalin fixed and paraffin embedded human skin tissue followed by peroxidase conjugation of the secondary antibody and DAB staining.This data demonstrates the use of p53 Antibody (T55) for immunohistochemistry. Clinical relevance has not been evaluated.

Formalin-fixed and paraffin-embedded human lung carcinoma tissue reacted with p53 Antibody (S315) (Cat.#TA324825), which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated.

MCF-7 cell extract was resolved by electrophoresis, transferred to nitrocellulose and probed with monoclonal anti-p53 (clone BP53-12) antibody. Proteins were visualized using a goat anti-mouse secondary conjugated to HRP and a chemiluminescence detection system.

ChIP assays were performed using human U2OS cells, treated with camptothecin, the ab against p53 and optimized PCR primer sets for QPCR. ChIP experiment was analysed. IgG (2 ug/IP) was used as negative IP control. QPCR was performed with primers for the p21 and GAS6 genes used as positive controls, and for GAPDH gene and the Sat2 satellite repeatas negative controls. Image shows the recovery, expressed as a % of input (the relative amount of IP'd DNA compared to input DNA after qPCR analysis).

ChIP was performed on sheared chromatin from 4 million U2OS cells using 1 ug of the ab against p53 as described above. The IP'd DNA was subsequently analysed on an Illumina HiSeq. The 51 bp tags were aligned to the human genome using the BWA algorithm. Image shows the peak distribution along the X-chromosome (fig 2A) and in 3 genomic regions of chromosome 6, 13 and 12, surrounding p21 (CDKN1A), GAS6 and MDM2, 3 known targets genes of p53 (fig 2B, C and D, respectively).

Immunofluorescent staining of COS7 cells using TP53 mouse monoclonal antibody (UM500049, green). Actin filaments were labeled with TRITC-phalloidin (red), and nuclear with DAPI (blue). The three-color overlay image is located at the bottom-right corner.

Immunofluorescent staining of HT-29 cells using TP53 mouse monoclonal antibody (UM500049, green). Actin filaments were labeled with TRITC-phalloidin (red), and nuclear with DAPI (blue). The three-color overlay image is located at the bottom-right corner.

Immunofluorescent staining of HT-29 cells using TP53 mouse monoclonal antibody (UM500053, green). Actin filaments were labeled with TRITC-phalloidin (red), and nuclear with DAPI (blue). The three-color overlay image is located at the bottom-right corner.

Immunofluorescent staining of COS7 cells using TP53 mouse monoclonal antibody (UM500053, green). Actin filaments were labeled with TRITC-phalloidin (red), and nuclear with DAPI (blue). The three-color overlay image is located at the bottom-right corner.

OriGene overexpression protein microarray chip was immunostained with UltraMAB anti-TP53 mouse monoclonal antibody (UM500053). The positive reactive proteins are highlighted with two red arrows in the enlarged subarray. All the positive controls spotted in this subarray are also labeled for clarification.

Immunohistochemical analysis of p53 staining in human colon cancer formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). The section was then incubated with the antibody at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

Immunofluorescent analysis of p53 staining in Hela cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with the primary antibody in 3% BSA-PBS and incubated overnight at 4 deg C in a hidified chamber. Cells were washed with PBST and incubated with a FITC-conjugated secondary antibody (green) in PBS at room temperature in the dark. DAPI was used to stain the cell nuclei (blue).